A novel human muscle cell model of Duchenne muscular dystrophy created by CRISPR/Cas9 and evaluation of antisense-mediated exon skipping

Shimo, Takenori; Hosoki, Kana; Nakatsuji, Yusuke; Yokota, Toshifumi; Obika, Satoshi

doi:10.1038/s10038-017-0400-0

Cited by 26 publications

(9 citation statements)

References 30 publications

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“…Accordingly, these limitations pose problems when evaluating the efficacy of exon skipping with AOs designed for patients, particularly multiple exon skipping using many AOs. For these issues, there exist some solutions such as the use of immortalized DMD muscle cell lines [122,123], fibroblast-converted muscle cells [81,124], DMD-patient-derived iPSC lines that can differentiate to muscle cells [125,126,127,128], muscle cell lines with artificial mutations [129], or cells loading a DMD construct mediated-reporter system [130].…”

Section: Patient-derived Cells and Humanized Animal Models For Tesmentioning

confidence: 99%

“…For the limitation concerning available mutations for study, a genome editing system can provide a solution to create new cell lines with artificial mutations that mimic natural mutations. Such genetically engineered cells have been reported with DMD iPSCs and a rhabdomyosarcoma cell line that are amenable to skipping exons 45–55 [59,129] and exons 3–9 [75]. In order to develop multiple exon skipping therapies, a variety of combination cocktails of AOs needs to be tested tailored to different deletion patterns.…”

Section: Patient-derived Cells and Humanized Animal Models For Tesmentioning

confidence: 99%

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Multiple Exon Skipping in the Duchenne Muscular Dystrophy Hot Spots: Prospects and Challenges

Echigoya

Lim

Nakamura

et al. 2018

JPM

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Duchenne muscular dystrophy (DMD), a fatal X-linked recessive disorder, is caused mostly by frame-disrupting, out-of-frame deletions in the dystrophin (DMD) gene. Antisense oligonucleotide-mediated exon skipping is a promising therapy for DMD. Exon skipping aims to convert out-of-frame mRNA to in-frame mRNA and induce the production of internally-deleted dystrophin as seen in the less severe Becker muscular dystrophy. Currently, multiple exon skipping has gained special interest as a new therapeutic modality for this approach. Previous retrospective database studies represented a potential therapeutic application of multiple exon skipping. Since then, public DMD databases have become more useful with an increase in patient registration and advances in molecular diagnosis. Here, we provide an update on DMD genotype-phenotype associations using a global DMD database and further provide the rationale for multiple exon skipping development, particularly for exons 45–55 skipping and an emerging therapeutic concept, exons 3–9 skipping. Importantly, this review highlights the potential of multiple exon skipping for enabling the production of functionally-corrected dystrophin and for treating symptomatic patients not only with out-of-frame deletions but also those with in-frame deletions. We will also discuss prospects and challenges in multiple exon skipping therapy, referring to recent progress in antisense chemistry and design, as well as disease models.

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Section: Patient-derived Cells and Humanized Animal Models For Tesmentioning

confidence: 99%

Section: Patient-derived Cells and Humanized Animal Models For Tesmentioning

confidence: 99%

Multiple Exon Skipping in the Duchenne Muscular Dystrophy Hot Spots: Prospects and Challenges

Echigoya

Lim

Nakamura

et al. 2018

JPM

Self Cite

View full text Add to dashboard Cite

show abstract

“…Various groups have gone on to develop other DMD animal models using CRISPR/Cas9; at present, the list includes mice with other Dmd mutations [59,82], rats [85], pigs [86], rabbits [87], and monkeys [88]. In vitro models of DMD have also been created using CRISPR/Cas9, by generating DMD mutations in rhabdomyosarcoma cells [89,90] or in hiPSCs [57]. We will not be describing the characteristics of these models here, but instead gladly invite the reader to consult the cited references for additional information.…”

Section: Dmd Studies In Vivomentioning

confidence: 99%

Applications of CRISPR/Cas9 for the Treatment of Duchenne Muscular Dystrophy

Lim

Yoon

Yokota

2018

JPM

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Duchenne muscular dystrophy (DMD) is a fatal X-linked recessive neuromuscular disease prevalent in 1 in 3500 to 5000 males worldwide. As a result of mutations that interrupt the reading frame of the dystrophin gene (DMD), DMD is characterized by a loss of dystrophin protein that leads to decreased muscle membrane integrity, which increases susceptibility to degeneration. CRISPR/Cas9 technology has garnered interest as an avenue for DMD therapy due to its potential for permanent exon skipping, which can restore the disrupted DMD reading frame in DMD and lead to dystrophin restoration. An RNA-guided DNA endonuclease system, CRISPR/Cas9 allows for the targeted editing of specific sequences in the genome. The efficacy and safety of CRISPR/Cas9 as a therapy for DMD has been evaluated by numerous studies in vitro and in vivo, with varying rates of success. Despite the potential of CRISPR/Cas9-mediated gene editing for the long-term treatment of DMD, its translation into the clinic is currently challenged by issues such as off-targeting, immune response activation, and sub-optimal in vivo delivery. Its nature as being mostly a personalized form of therapy also limits applicability to DMD patients, who exhibit a wide spectrum of mutations. This review summarizes the various CRISPR/Cas9 strategies that have been tested in vitro and in vivo for the treatment of DMD. Perspectives on the approach will be provided, and the challenges faced by CRISPR/Cas9 in its road to the clinic will be briefly discussed.

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“…Various groups have gone on to develop other DMD animal models using CRISPR/Cas9; at present, the list includes mice with other Dmd mutations [59,81], rats [82], pigs [83], rabbits [84], and monkeys [85]. In vitro models of DMD have also been created using CRISPR/Cas9, by generating DMD mutations in rhabdomyosarcoma cells [86,87] or in hiPSCs [57]. We will not be describing the characteristics of these models here, but instead gladly invite the reader to consult the cited references for additional information.…”

Section: Crispr/cas9 For the Generation Of Dmd Animal/cell Modelsmentioning

confidence: 99%

Methods of CRISPR/Cas9 Exon Skipping for Duchenne Muscular Dystrophy

Krq¹,

Yoon²,

Yokota³

2018

Preprint

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Duchenne muscular dystrophy (DMD) is a fatal X-linked recessive neuromuscular disease prevalent in 1 in 3500 to 5000 males worldwide. As a result of mutations that interrupt the reading frame of the dystrophin gene (DMD), DMD is characterized by a loss of dystrophin protein which leads to decreased muscle membrane integrity, which increases susceptibility to degeneration. CRISPR/Cas9 technology has garnered interest as an avenue for DMD therapy due to its potential for permanent exon skipping, which can restore the disrupted DMD reading frame in DMD and lead to dystrophin restoration. An RNA-guided DNA endonuclease system, CRISPR/Cas9 allows for the targeted editing of specific sequences in the genome. The efficacy and safety of CRISPR/Cas9 as a therapy for DMD has been evaluated by numerous studies in vitro and in vivo, with varying rates of success. Despite the potential of CRISPR/Cas9-mediated gene editing for the long-term treatment of DMD, its translation into the clinic is currently challenged by issues such as off-targeting, immune response activation, and sub-optimal in vivo delivery. Its nature as being mostly a personalized form of therapy also limits applicability to DMD patients, who exhibit a wide spectrum of mutations. This review summarizes the various CRISPR/Cas9 strategies that have been tested in vitro and in vivo for the treatment of DMD. Perspectives on the approach will be provided, and the challenges faced by CRISPR/Cas9 in its road to the clinic will be briefly discussed.

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A novel human muscle cell model of Duchenne muscular dystrophy created by CRISPR/Cas9 and evaluation of antisense-mediated exon skipping

Cited by 26 publications

References 30 publications

Multiple Exon Skipping in the Duchenne Muscular Dystrophy Hot Spots: Prospects and Challenges

Multiple Exon Skipping in the Duchenne Muscular Dystrophy Hot Spots: Prospects and Challenges

Applications of CRISPR/Cas9 for the Treatment of Duchenne Muscular Dystrophy

Methods of CRISPR/Cas9 Exon Skipping for Duchenne Muscular Dystrophy

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